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目的研究牛大力多糖对脂多糖(LPS)诱导的小鼠巨噬细胞RAW264.7细胞分泌炎症因子的影响,并观察核转录因子-κB p65(NF-κB p65)和抑制性卡巴蛋白-α(IκB-α)表达的变化。方法选用小鼠巨噬细胞RAW264.7,随机分为空白对照组,LPS模型组,牛大力多糖低、中、高剂量组(10,100,1000 ng·m L~(-1)),加药孵育2 h后,再加入10μg·m L~(-1)LPS至培养板中继续培养10 h,ELISA法测定各组细胞炎性因子白细胞介素1(IL-1)、白细胞介素6(IL-6)、肿瘤坏死因子α(TNF-α)的含量,Western Blot法检测IκB-α蛋白以及NF-κB p65蛋白的表达水平。结果与LPS模型组比较,牛大力多糖高、中、低剂量组均能不同程度地抑制LPS诱导的3种炎性因子IL-1、IL-6、TNF-α的释放(P<0.05,P<0.01),提高IκB-α蛋白表达(P<0.05,P<0.01),降低NF-κB p65蛋白的表达(P<0.05,P<0.01),且牛大力多糖对上述指标的作用强度与剂量呈正相关,各组间比较差异均有统计学意义(P<0.05,P<0.01)。结论牛大力多糖可通过增加IκB-α蛋白和抑制NF-κB p65蛋白的表达,从而降低炎症因子IL-1、IL-6、TNF-α的释放,起到抗炎作用。
Objective To investigate the effects of bovine polysaccharides on the secretion of inflammatory cytokines in RAW 264.7 cells induced by lipopolysaccharide (LPS) and to observe the effects of nuclear factor-κB p65 (NF-κB p65) and inhibitory kappa-protein alpha ( IκB-α) expression changes. Methods Mouse macrophage RAW264.7 cells were randomly divided into blank control group, LPS model group, low, medium and high doses of bovine polysaccharide (10, 100, 1000 ng · m L -1) After 2 h, 10μg · m L -1 LPS was added to the culture plate for further cultivation for 10 h. The levels of inflammatory cytokines IL-1, IL-6 -6) and tumor necrosis factor-α (TNF-α) were detected by ELISA. The expression of IκB-α protein and NF-κB p65 protein were detected by Western Blot. Results Compared with LPS model group, high, medium and low doses of LPS could inhibit the release of IL-1, IL-6 and TNF-α induced by LPS to some extent (P <0.05, P (P <0.05, P <0.01), and increased the expression of IκB-α protein (P <0.05, P <0.01) and decreased the expression of NF-κB p65 protein (P <0.05, P <0.01). There was a positive correlation between the two groups. Conclusion Dali polysaccharides can reduce the release of inflammatory cytokines IL-1, IL-6 and TNF-α by increasing the expression of IκB-α and inhibiting the expression of NF-κB p65 protein.